Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.
Bees/genetics* ; Animals ; Larva/metabolism* ; RNA, Circular/genetics* ; RNA, Small Interfering/genetics* ; MicroRNAs/genetics*

As the most efficient perception system in nature, the perception mechanism of the insect (such as honeybee) antennae is the key to imitating the high-performance sensor technology. An automated experimental device suitable for collecting electrical signals (including antenna reaction time information) of antennae was developed, in response to the problems of the non-standardized experimental process, interference of manual operation, and low efficiency in the study of antenna perception mechanism. Firstly, aiming at the automatic identification and location of insect heads in experiments, the image templates of insect head contour features were established. Insect heads were template-matched based on the Hausdorff method. Then, for the angle deviation of the insect heads relative to the standard detection position, a method that calculates the angle of the insect head mid-axis based on the minimum external rectangle of the long axis was proposed. Eventually, the electrical signals generated by the antennae in contact with the reagents were collected by the electrical signal acquisition device. Honeybees were used as the research object in this study. The experimental results showed that the accuracy of template matching could reach 95.3% to locate the bee head quickly, and the deviation angle of the bee head was less than 1°. The distance between antennae and experimental reagents could meet the requirements of antennae perception experiments. The parameters, such as the contact reaction time of honeybee antennae to sucrose solution, were consistent with the results of the manual experiment. The system collects effectively antenna contact signals in an undisturbed state and realizes the standardization of experiments on antenna perception mechanisms, which provides an experimental method and device for studying and analyzing the reaction time of the antenna involved in biological antenna perception mechanisms.
Animals ; Arthropod Antennae ; Bees

Aims: Several cockroach and ant species have been revealed to infest households with inadequate insect control and food storage practices. These household insects harbor countless bacteria species of public health, agricultural and industrial importance. Many studies have reported disease-causing bacteria from both cockroaches and ant’s species collected from hospitals and residential areas. The aim of this study was to characterize the culturable bacterial communities of two common household insects, big headed ants (Pheidole rugaticeps) and American cockroaches (Periplaneta americana) using 16S rRNA genes sequencing. Methodology and results: A total of 64 bacterial sequences were obtained from P. rugaticeps (48.44%) and P. americana (51.56%) and Firmicutes was the most dominant phylum from both insect species. Bacillus was the most dominant bacterial genus from both cockroach and ant samples. Other important genera isolated were Pseudomonas and Stenotrophomonas which have previously been suggested to have members that are of biotechnological importance. Food poisoning bacterial species, B. cereus and other bacterial strains such as B. subtilis, Acinetobacter baumannii, Burkholderia cepacia, P. aeruginosa, Staphylococcus epidermidis, Serratia marcescens and S. pseudintermedius with the history of human infections were isolated from some of the insect’s specimens. Conclusion, significance and impact of study Thus, these household insect pests harbor bacterial species known to cause diseases of serious public health importance that needs serious attentions. Similarly, the insects harbor other bacteria species that may provide opportunities for biotechnological exploration.
Ants ; Periplaneta ; Public health

Aims: Symbiotic bee fungus associated with the stingless bee larval food has been revealed to play a major role in performing a mutual relationship with the host. The fungus is believed capable to produce crucial nutrients that are required for larval pupation. However, detailed information on the fungus identification isolated from the larval food of our native Indo-Malaya stingless bee, Heterotrigona itama (Hymenoptera: Apidae), is poorly understood. Hence, this study was conducted to determine the effect of fungus isolated from the stingless bee larval food on the survival of H. itama larvae and identify the isolated fungus using both morphological and molecular analyses. Methodology and results: Elisa plates designed with F-bottom type were used as artificial brood cells in a controlled condition. The eggs transferred to Elisa plates were kept in the incubator with 75%-100% humidity and 25 ± 2.5 °C of temperature. This study carried out in three different treatments: (1) microbes-free larval food, (2) fresh larval food and (3) microbes-free larval food with fungus supplement. Results showed that the survival of H. itama larvae depends on the presence of beneficial fungus with the highest survival rate (Treatment 2 = 85.71%). The absence of the beneficial fungus in the fluid food of H. itama showed the lowest survival rate (Treatment 1 = 37.14%). Microscopy analysis showed that the fungus had various forms, including unconjugated asci, round-shaped ascospore and pseudohyphae. The molecular characterisation of the isolated fungi was performed using 23S rRNA gene sequencing using universal primers ITS1 and ITS4. DNA barcoding of three isolated fungi confirmed all isolated fungi matched Panus lecomtei (Basidiomycetes; Poriales; Polyporaceae) with 99.70% of similarity. Conclusion, significance and impact of study This study provides information on the importance of whitish fungus in appearance that existed in the brood cell, which is proposed to be the crucial component of in-vitro stingless bee queen rearing protocol.
Bees--microbiology

Six new (1-6) and seven known depsidones (7-13) were isolated from the culture of an ant (Monomorium chinensis)-derived fungus Spiromastix sp. MY-1. Their structures were elucidated by extensive spectroscopic analysis including high resolution MS, 1D and 2D NMR data. The new bromide depsidones were obtained through supplementing potassium bromide in the fermentation medium of Spiromastix sp. MY-1. All isolated compounds showed various bioactivities against the tested phytopathogenic bacteria. Particularly, new bromide compound 4, named spiromastixone S, exhibited the strongest activity against Xanthomonas oryzae pv. oryzae with a MIC value of 5.2 μmol·^-1.
Animals ; Anti-Bacterial Agents ; Ants ; Bromides ; Depsides ; Fungi ; Lactones ; Microbial Sensitivity Tests ; Molecular Structure

Two terpenes, 3-keto-tirucalla-8,24-dien-21-oic acid(KTDA) and 2-methoxy-5-acetoxy-furanogermacr-1(10)-en-6-one(FSA), are isolated from Olibanum and Myrrha respectively, which are characterized by high yield and easy crystallization during the preparation. The present study explored the regulatory targets and anti-inflammatory mechanism of KTDA and FSA based on network pharmacology and cell viability assay. First, the drug-likeness of KTDA and FSA was predicted by Swiss ADME. The target prediction of active components was carried out by Swiss Target Prediction and Pharmmapper. TTD, Drug Bank, and Gene Cards were searched for inflammation-related target genes of KTDA and FSA. Protein-protein interaction(PPI) analysis was performed on the inflammatory targets of KTDA and FSA by STRING, and Cytoscape was used to conduct topological analysis of the interaction results and construct the PPI network. GO function and KEGG pathway enrichment analyses of inflammatory targets of KTDA and FSA were carried out by DAVID, and a " component-target-pathway" network was constructed. Finally, lipopolysaccharide(LPS)-induced RAW264. 7 cells were treated with KTDA and FSA at different concentrations, and nitric oxide(NO) concentration and protein and m RNA expression levels were detected. The results showed that both KTDA and FSA showed good drug-likeness. A total of 157 and 142 inflammation-related targets of KTDA and FSA were screened out. PPI network analysis showed that MAPK1, AKT1, MAPK8, PIK3 CA,PIK3 R1, EGFR, etc. might be the key proteins for the anti-inflammatory effect. PI3 K/AKT and MAPK signaling pathways were obtained by KEGG and GO-BP enrichment. Cell experiment results showed that KTDA and FSA could exert anti-inflammatory effects by inhibiting NO production, reducing the phosphorylation levels of JNK, p38, and AKT proteins, and down-regulating the m RNA expression of interleukin(IL)-1β and IL-6. Meanwhile, FSA could also inhibit ERK phosphorylation. The results indicated that KTDA and FSA had significant anti-inflammatory activity, which provided a scientific basis and important support for the further research,development, and utilization of Olibanum and Myrrha.
Animals ; Ants ; Drugs, Chinese Herbal/pharmacology* ; Frankincense ; Lipopolysaccharides ; Molecular Docking Simulation ; Network Pharmacology

bees (Apis mellifera), and then, within 10 min, amplified the target cDNA by ultra-rapid qPCR. The PCR products were hybridized to a DNA-chip for confirmation of target gene specificity. The results of this study suggest that our method might be a useful tool for detecting DWV, as well as for the diagnosis of RNA virus-mediated diseases on-site.]]>
Beekeeping ; Bees ; Diagnosis ; DNA, Complementary ; Methods ; Polymerase Chain Reaction ; RNA ; RNA, Viral ; Sensitivity and Specificity

PURPOSE: To report a patient stung by a bee, who was diagnosed with sterile endopthalmitis and another patient diagnosed with optic neuritis, with decreasing visual acuity, after refined bee venom injection around the orbital tissue. CASE SUMMARY: A 82-year-old female visited our hospital for decreased visual acuity in the right eye and ocular pain due to a bee sting. The bee sting penetrated the sclera into the vitreous. In the anterior segment, severe cornea edema and anterior chamber cells were seen. Using ultrasonography, inflammation was seen around the intravitreal area. After 3 months, intravitreal inflammation regressed but the patient's visual acuity was light perception negative, and corneal opacity, neovascularization, and phthisis bulbi were detected. A 55-year-old male visited our hospital for ocular pain in the right eye and decreasing visual acuity after refined bee venom injection around the orbital tissue. The best-corrected visual acuity in the right eye was 15/100, there was moderate injection on the conjunctiva. A relative afferent pupillary defect, abnormal color vision test results, and a defect in the visual field test were observed. There was no pain during external ocular movement, and other general blood tests, and a brain MRI were normal. Based on these symptoms, methylprednisolone megatherapy was started for treatment of optic neuritis. After treatment, visual acuity of the right eye was 9/10 and all other clinical optic neuritis symptoms regressed. CONCLUSIONS: Based on these two cases, ocular toxicity from bee venom could result from both direct and indirect courses. Treatment using refined bee venom might be harmful, and caution is recommended in its use.
Aged, 80 and over ; Anterior Chamber ; Bee Venoms ; Bees ; Bites and Stings ; Brain ; Color Vision ; Conjunctiva ; Cornea ; Corneal Opacity ; Edema ; Female ; Hematologic Tests ; Humans ; Inflammation ; Magnetic Resonance Imaging ; Male ; Methylprednisolone ; Middle Aged ; Optic Neuritis ; Orbit ; Pupil Disorders ; Sclera ; Ultrasonography ; Visual Acuity ; Visual Field Tests

Anaphylaxis usually develop immediately after wasp sting, but may develop even after few days later. Neurological complications after stings are uncommon, although several cases have been reported involving central and/or peripheral nervous system. Although wasp sting-induced encephalitis has been rarely reported, all reported cases showed mental change and severe neurological deterioration. Herein, we report an atypical case who showed biphasic anaphylaxis and delayed-onset cerebellar ataxia following a wasp sting, characterized by mild cerebellar ataxia and excellent response to corticosteroids.
Adrenal Cortex Hormones ; Anaphylaxis ; Bites and Stings ; Cerebellar Ataxia ; Encephalitis ; Peripheral Nervous System ; Wasps

BACKGROUND: Understanding the epidemiology of anaphylaxis is imperative for appropriate diagnosis and treatment, but the prevalence reportedly varies and only a few studies have compared the clinical features of anaphylaxis with the underlying causes in Korea. OBJECTIVE: This study aimed to investigate the etiology and clinical features of anaphylaxis. METHODS: We retrospectively reviewed the medical records of 319 anaphylaxis patients who visited our emergency room and extracted information on the causes, clinical characteristics, and subsequent outpatient visits. RESULTS: Food, drugs, and environmental factors were common causes of anaphylaxis. Statistically significant differences (p<0.001) were observed between children (<18 years of age) and adults (≥18 years of age), with food and drugs identified as the most common causes, respectively. Clinical characteristics of the patients were compared according to the common causes of anaphylaxis. Cutaneous symptoms were observed more frequently in food-induced cases (n=137, 95.1%) compared to drug-induced (n=73, 77.7%) and bee sting-induced (n=18, 78.3%) cases (p<0.001), whereas neurological symptoms were observed more frequently in drug-induced (n=37, 39.4%) and bee sting-induced (n=11, 47.8%) cases than in food-induced (n=18, 12.5%) cases (p<0.01). Drug-induced (n=44, 46.8%) and bee sting-induced (n=11, 47.8%) cases were more severe than food-induced cases (n=32, 22.2%). Out of the 319 assessed patients, only 25, 14, and 14 patients were referred to the pediatrics, allergy, and dermatology departments, respectively, after visiting the emergency room. CONCLUSION: Clinical characteristics of anaphylactic patients differed according to the underlying cause, but these findings are presumably influenced by factors determining the severity of anaphylaxis. We found that subsequent follow-up care in other departments to identify the cause of anaphylaxis was inadequate for most patients.
Adult ; Anaphylaxis ; Bees ; Child ; Dermatology ; Diagnosis ; Emergencies ; Emergency Service, Hospital ; Epidemiology ; Follow-Up Studies ; Humans ; Hypersensitivity ; Korea ; Medical Records ; Outpatients ; Pediatrics ; Prevalence ; Retrospective Studies ; Tertiary Care Centers